Target-serine-threonine phosophatase 2C Toxoplasma Gondii

ORGANISM: Toxoplasma gondii ME49

GENE of INTEREST: TGME49_031850 (serine-threonine phosophatase 2C)


BACKGROUND/ DISEASE:

IMAGE:

NCBI GENE # or RefSeq#: 7898899


EC# OF ENZYME: 3.1.3.16


CDS GENE SEQUENCE:

        1 atgaagtcct ctgctgaaat taggcggacc atggatgtcc ctcctaccat tcatgtacct
       61 ctccctccaa cgtcgtatcc cgctttcgat gctgcgatct tcacagacat cggtgggcgc
      121 aagcatcagg aagataggtt cactctctgt ccgcagctcg ttcccggccg agacgactgc
      181 gccttcttcg gtgtcttcga tggcactgtt ggagatttcg ccagcgaaaa tgtgaaggat
      241 cttgttgttc cacagttgat ttcctcgccc gcgtggcagg aggtgactga gatgctgaga
      301 tcagacgtac ccgccactga ggtggacgag aagctccctc agttgcttga tcaggcagtc
      361 gatgacatgt acaagaacgc agacaacgaa cttgtgaaga tgtgcgagca gcttaacaaa
      421 gactacgcga gcagcacctc cgtcacggca gtcttggcca aaggcttcgt ggctgttggt
      481 catctgggcg acagccgcat cgctatggga gtcgagacgc caaacgggtt gaactgcgag
      541 ttcttgaccg ttgaccacaa gccggatatg ccacatgaga aactgcgcat catgcgcaat
      601 ggaggcagtg ttgagtatct ccacaaccac aacaacaaac cgttcattcg aggtggtgac
      661 ttctcgttcc ggaagtcgcg cggagagcag ccgatgcagc tccagtactc ccgagctttc
      721 ggcgggaagg acctgaagat gtacggtttg agcaaccaac ccgacgtacg cgttgtccgc
      781 gtcacgcctc aacaccgagt gatgattctc gcgactgatg gcttgtggga cgtcatgtct
      841 gcggcgcaag ctgtagagat cgctatgcag gcacgacagg aaggaaggaa cccagcgcag
      901 gcgctggtgg agatgaccct cgctgagcag cagagccgca accaaagtgc agacaacatt
      961 actgcgatga cagtgttctt caagaagact gattag</span>

AMINO ACID SEQUENCE:

SLTMDVPPTIHVPLPPTSYPAFDAAIFTDIGGRKHQEDRFTLCPQLVPGRDDCAFFGVFDGTVGDFASENVKD

LVVPQLISSPAWQEVTEMLRSDVPATEVDEKLPQLLDQAVDDMYKNADNELVKMCEQLNKDYASSTSVTAVLA

KGFVAVGHLGDSRIAMGVETPNGLNCEFLTVDHKPDMPHEKLRIMRNGGSVEYLHNHNNKPFIRGGDFSFRKS

RGEQPMQLQYSRAFGGKDLKMYGLSNQPDVRVVRVTPQHRVMILATDGLWDVMSAAQAVEIAMQARQEGRNPA

QALVEMTLAEQQSRNQSADNTAMTVFFKKTD


PDB:
http://www.pdb.org/pdb/explore/explore.do?structureId=2I44

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Primer Design - Larry

Upstream
5’ TACTTCCAATCCATGAAGTCCTCTGCTGAAAT 3’

Downstream
5’ TATCCACCTTTACTGCTAATCAGTCTTCTTGAAGA 3’
Downstream Reverse Complement
5’ TCTTCAAGAAGACTGATTAGCAGTAAAGGTGGATA 3’

1 ATGAAGTCCT CTGCTGAAAT TAGGCGGACC ATGGATGTCC CTCCTACCAT TCATGTACCT
61 CTCCCTCCAA CGTCGTATCC CGCTTTCGAT GCTGCGATCT TCACAGACAT CGGTGGGCGC
121 AAGCATCAGG AAGATAGGTT CACTCTCTGT CCGCAGCTCG TTCCCGGCCG AGACGACTGC
181 GCCTTCTTCG GTGTCTTCGA TGGCACTGTT GGAGATTTCG CCAGCGAAAA TGTGAAGGAT
241 CTTGTTGTTC CACAGTTGAT TTCCTCGCCC GCGTGGCAGG AGGTGACTGA GATGCTGAGA
301 TCAGACGTAC CCGCCACTGA GGTGGACGAG AAGCTCCCTC AGTTGCTTGA TCAGGCAGTC
361 GATGACATGT ACAAGAACGC AGACAACGAA CTTGTGAAGA TGTGCGAGCA GCTTAACAAA
421 GACTACGCGA GCAGCACCTC CGTCACGGCA GTCTTGGCCA AAGGCTTCGT GGCTGTTGGT
481 CATCTGGGCG ACAGCCGCAT CGCTATGGGA GTCGAGACGC CAAACGGGTT GAACTGCGAG
541 TTCTTGACCG TTGACCACAA GCCGGATATG CCACATGAGA AACTGCGCAT CATGCGCAAT
601 GGAGGCAGTG TTGAGTATCT CCACAACCAC AACAACAAAC CGTTCATTCG AGGTGGTGAC
661 TTCTCGTTCC GGAAGTCGCG CGGAGAGCAG CCGATGCAGC TCCAGTACTC CCGAGCTTTC
721 GGCGGGAAGG ACCTGAAGAT GTACGGTTTG AGCAACCAAC CCGACGTACG CGTTGTCCGC
781 GTCACGCCTC AACACCGAGT GATGATTCTC GCGACTGATG GCTTGTGGGA CGTCATGTCT
841 GCGGCGCAAG CTGTAGAGAT CGCTATGCAG GCACGACAGG AAGGAAGGAA CCCAGCGCAG
901 GCGCTGGTGG AGATGACCCT CGCTGAGCAG CAGAGCCGCA ACCAAAGTGC AGACAACATT
961 ACTGCGATGA CAGTGTTCTT CAAGAAGACT GATTAG

Melting Temp

	GC Content	0mM Mg++	1.5mM Mg++	2mM Mg++	4mM Mg++	6mM Mg++
Forward	40.0 %	52.0 ºC	59.0 ºC	59.8 ºC	61.3 ºC	62.0 ºC
Reverse	35.0 %	47.0 ºC	54.2 ºC	55.0 ºC	56.5 ºC	57.2 ºC

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Primer Design - Justin

Forward Primer:
5’ TACTTCCAATCCATGGATGTCCCGCCGACCATC 3’ 33 bp
GC Content 54.5%
0 mM Mg2+ Tm 67.0 oC 1.5 mM Mg2+ Tm 73.5 oC 2 mM Mg2+ Tm 73.9 oC
4 mM Mg2+ Tm 74.8 oC 6 mM Mg2+ Tm 75.1 oC


Reverse Primer:
5’ TTCAAAAAAACCGACTAACAGTAAAGGTGGATA 3’

Reverse Complemented:
5’ TATCCACCTTTACTGTTAGTCGGTTTTTTTGAA 3’ 33 bGC Content 33.3%
0 mM Mg2+ Tm 58.7 oC 1.5 mM Mg2+ Tm 66.8 oC 2 mM Mg2+ Tm 67.3 oC
4 mM Mg2+ Tm 68.5 oC 6 mM Mg2+ Tm 69.0 oC

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Primer Design - Jericho

OUTPUT FROM DNA WORKS

TCTCTGACCATGGATGTACCACCTACCATTCACGTTCCTCTGCCGCCGACGTCTTACCCG
GCGTTCGACGCGGCGATCTTCACCGACATCGGTGGCCGTAAACACCAGGAAGACCGCTTC
ACCCTGTGCCCGCAGCTGGTTCCGGGTCGTGACGACTGCGCGTTCTTCGGTGTTTTCGAT
GGTACCGTTGGTGACTTCGCGTCTGAAAACGTTAAGGACCTGGTTGTTCCTCAGCTCATC
TCTTCTCCGGCGTGGCAGGAAGTGACCGAAATGCTGCGTTCTGATGTTCCAGCCACTGAG
GTTGATGAGAAACTGCCGCAACTGCTGGACCAGGCGGTAGACGACATGTACAAAAACGCG
GACAACGAGCTGGTGAAAATGTGCGAACAGCTGAACAAAGACTACGCGTCTTCTACCTCT
GTTACCGCCGTTCTGGCGAAAGGTTTCGTTGCGGTTGGTCACCTGGGTGACTCTCGTATT
GCGATGGGTGTTGAAACCCCGAACGGTCTCAACTGCGAATTCCTGACCGTTGACCACAAA
CCGGACATGCCGCACGAAAAACTGCGTATTATGCGTAACGGTGGTTCTGTTGAATACCTG
CACAACCACAACAACAAGCCGTTCATCCGTGGTGGCGACTTCTCTTTCCGTAAATCTCGT
GGTGAACAGCCGATGCAGCTCCAGTATTCTCGTGCGTTCGGTGGTAAAGACCTCAAAATG
TACGGTCTGTCTAACCAGCCGGACGTTCGTGTTGTGCGTGTTACTCCGCAGCACCGTGTT
ATGATCCTGGCGACCGATGGTCTGTGGGACGTTATGTCTGCGGCGCAGGCCGTTGAGATT
GCCATGCAAGCACGTCAAGAAGGTCGTAATCCGGCGCAAGCGCTCGTCGAAATGACCCTG
GCGGAACAGCAGTCTCGTAACCAGAGCGCCGACAACACCGCGATGACGGTTTTCTTCAAA
AAAACCGACTAA

Melting Temperatures

Upstream:
5’ TACTTCCAATCCATGATGGATGTACCACCTACCATT 3’ 36 bp GC Content 41.7 %

0 mM Mg2+ Tm 63.0 ºC
1.5 mM Mg2+ Tm 70.5 ºC
2 mM Mg2+ Tm 71.0 ºC
4 mM Mg2+ Tm 72.0 ºC
6 mM Mg2+ Tm 72.4 ºC

Downstream:
5’ TTCTTCAAAAAAACCGACTAACAGTAAAGGTGGATA 3’

Reverse Complement:
5’ TATCCACCTTTACTGTTAGTCGGTTTTTTTGAAGAA 3’ 36 bp GC Content 33.3 %

0 mM Mg2+ Tm 59.9 ºC
1.5 mM Mg2+ Tm 68.1 ºC
2 mM Mg2+ Tm 68.6 ºC
4 mM Mg2+ Tm 69.7 ºC
6 mM Mg2+ Tm 70.2 ºC